127 results on '"Hira"'
Search Results
102. HIRA, a DiGeorge Syndrome Candidate Gene, Confers Proper Chromatin Accessibility on HSCs and Supports All Stages of Hematopoiesis.
- Author
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Chen, Chao, Sun, Ming-an, Warzecha, Claude, Bachu, Mahesh, Dey, Anup, Wu, Tiyun, Adams, Peter D, Macfarlan, Todd, Love, Paul, and Ozato, Keiko
- Abstract
HIRA is a histone chaperone that deposits the histone variant H3.3 in transcriptionally active genes. In DiGeorge syndromes, a DNA stretch encompassing HIRA is deleted. The syndromes manifest varied abnormalities, including immunodeficiency and thrombocytopenia. HIRA is essential in mice, as total knockout (KO) results in early embryonic death. However, the role of HIRA in hematopoiesis is poorly understood. We investigate hematopoietic cell-specific Hira deletion in mice and show that it dramatically reduces bone marrow hematopoietic stem cells (HSCs), resulting in anemia, thrombocytopenia, and lymphocytopenia. In contrast, fetal hematopoiesis is normal in Hira-KO mice, although fetal HSCs lack the reconstitution capacity. Transcriptome analysis reveals that HIRA is required for expression of many transcription factors and signaling molecules critical for HSCs. ATAC-seq analysis demonstrates that HIRA establishes HSC-specific DNA accessibility, including the SPIB/PU.1 sites. Together, HIRA provides a chromatin environment essential for HSCs, thereby steering their development and survival. • HIRA confers chromatin accessibility that defines the HSC transcriptome • Long-term HSCs in adult Hira KO incur increased DNA damage and apoptosis • Hira KO has some similarity to DiGeorge syndromes, which lack the HIRA locus • Fetal hematopoiesis is spared in Hira-KO mice Chen et al. show that the histone H3.3 chaperone HIRA directs development and survival of adult hematopoietic stem cells (HSCs). Hira deletion impairs development of all lineages of hematopoiesis. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
103. Dynamic Incorporation of Histone H3 Variants into Chromatin Is Essential for Acquisition of Aggressive Traits and Metastatic Colonization.
- Author
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Gomes, Ana P., Ilter, Didem, Low, Vivien, Rosenzweig, Adam, Shen, Zih-Jie, Schild, Tanya, Rivas, Martin A., Er, Ekrem E., McNally, Dylan R., Mutvei, Anders P., Han, Julie, Ou, Yi-Hung, Cavaliere, Paola, Mullarky, Edouard, Nagiec, Michal, Shin, Sejeong, Yoon, Sang-Oh, Dephoure, Noah, Massagué, Joan, and Melnick, Ari M.
- Subjects
- *
HISTONES , *CHROMATIN , *TRANSCRIPTION factors , *CANCER invasiveness , *COLONIZATION - Abstract
Metastasis is the leading cause of cancer mortality. Chromatin remodeling provides the foundation for the cellular reprogramming necessary to drive metastasis. However, little is known about the nature of this remodeling and its regulation. Here, we show that metastasis-inducing pathways regulate histone chaperones to reduce canonical histone incorporation into chromatin, triggering deposition of H3.3 variant at the promoters of poor-prognosis genes and metastasis-inducing transcription factors. This specific incorporation of H3.3 into chromatin is both necessary and sufficient for the induction of aggressive traits that allow for metastasis formation. Together, our data clearly show incorporation of histone variant H3.3 into chromatin as a major regulator of cell fate during tumorigenesis, and histone chaperones as valuable therapeutic targets for invasive carcinomas. • Metastasis inducers lead to a decline in CAF-1 suppressing canonical H3 incorporation • EMT and metastatic colonization occur as a function of CAF-1 levels • Histone H3.3 variant is essential for tumor progression and aggressive phenotypes • HIRA-mediated H3.3 gap filling induces a pro-metastatic transcriptional reprogramming Gomes et al. reveal metastatic stimuli reduce histone H3.1/H3.2 deposition on chromatin by suppressing the CAF-1 complex in breast and NSCLC cancer cells, leading to increased incorporation of non-canonical histone H3.3, which in turn induces chromatin remodeling and expression of metastatic genes. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
104. The impact of the HIRA histone chaperone upon global nucleosome architecture
- Author
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Konrad Paszkiewicz, Nicholas A. Kent, Karen M Moore, Simon K. Whitehall, and Csenge Gal
- Subjects
Nucleosome organization ,Nucleosome assembly ,Euchromatin ,HIRA ,nucleosome assembly ,Histones ,03 medical and health sciences ,0302 clinical medicine ,Schizosaccharomyces ,Nucleosome ,Promoter Regions, Genetic ,Molecular Biology ,030304 developmental biology ,Genetics ,0303 health sciences ,biology ,heterochromatin ,Cell Biology ,Chromatin Assembly and Disassembly ,biology.organism_classification ,Chromatin ,Nucleosomes ,Cell biology ,Histone ,histone chaperone ,Chromatosome ,biology.protein ,RNA Polymerase II ,Schizosaccharomyces pombe Proteins ,030217 neurology & neurosurgery ,Transcription Factors ,Reports ,S. pombe ,Developmental Biology - Abstract
HIRA is an evolutionarily conserved histone chaperone that mediates replication-independent nucleosome assembly and is important for a variety of processes such as cell cycle progression, development, and senescence. Here we have used a chromatin sequencing approach to determine the genome-wide contribution of HIRA to nucleosome organization in Schizosaccharomyces pombe. Cells lacking HIRA experience a global reduction in nucleosome occupancy at gene sequences, consistent with the proposed role for HIRA in chromatin reassembly behind elongating RNA polymerase II. In addition, we find that at its target promoters, HIRA commonly maintains the full occupancy of the −1 nucleosome. HIRA does not affect global chromatin structure at replication origins or in rDNA repeats but is required for nucleosome occupancy in silent regions of the genome. Nucleosome organization associated with the heterochromatic (dg-dh) repeats located at the centromere is perturbed by loss of HIRA function and furthermore HIRA is required for normal nucleosome occupancy at Tf2 LTR retrotransposons. Overall, our data indicate that HIRA plays an important role in maintaining nucleosome architecture at both euchromatic and heterochromatic loci.
- Published
- 2015
- Full Text
- View/download PDF
105. Notes about Censorship and Self-Censorship in the Biography of the Prophet Muḥammad
- Author
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Michael Lecker
- Subjects
Medina ,Muḥammad ,BP1-253 ,Ḥīra ,hadiz ,History of Civilization ,ʽabbāsíes ,CB3-482 ,Sīra ,censura ,Islam ,omeyas ,biografía - Abstract
The study of the medieval literary output about Muḥammad’s life should go hand in hand with the study of his history, for which we have rich evidence in a variety of sources. Ibn Isḥāq’s biography of Muḥammad and its epitome by Ibn Hishām were products of their time. A case of self-censorship applied by one of Ibn Isḥāq’s informants and two cases of censorship applied by Ibn Hishām, who omitted many of his predecessor’s materials, contribute to a better understanding of the social and political context of the biography., El estudio de la producción literaria medieval sobre la vida de Muḥammad debe ir de la mano del estudio de su historia, empresa para la que disponemos de rica información en una variedad de fuentes. La biografía de Muḥammad por Ibn Isḥāq y su epítome por Ibn Hišām fueron productos de su época. Un caso de auto-censura aplicado por uno de los informantes de Ibn Isḥāq y dos casos de censura aplicados por Ibn Hišām, quien omitió muchos de los materiales de su predecesor, contribuyen a una mejor comprensión del contexto social y político de la biografía del Profeta.
- Published
- 2014
- Full Text
- View/download PDF
106. Development and Validation of Quantitative PCR Assays for DNA-Based Newborn Screening of 22q11.2 Deletion Syndrome, Spinal Muscular Atrophy, Severe Combined Immunodeficiency and Congenital Cytomegalovirus Infection
- Author
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Theriault, Mylene A.
- Subjects
T-cell receptor excision circle ,congenital cytomegalovirus ,newborn screening ,TaqMan ,relative quantification ,UL54 ,copy number variation ,CRKL ,TBX1 ,HIRA ,severe combined immunodeficiency ,COMT ,dried blood spot ,OpenArray ,multiplex ,qPCR ,US17 ,22q11.2 deletion syndrome ,UFD1L ,UL55 ,contiguous gene deletion ,SMN1 ,spinal muscular atrophy ,SMN2 - Abstract
The development of new high throughput technologies able to multiplex disease biomarkers as well as advances in medical treatments has lead to the recent expansion of the newborn screening panel to include DNA-based targets. Four rare disorders; deletion 22q11.2 syndrome and Spinal Muscular Atrophy (SMA), Severe Combined Immunodeficiency (SCID) and Congenital Cytomegalovirus (CMV), are potential candidates for inclusion to the newborn screening panel within the next few years. The major focus of this study was to determine whether 5’-hydrolysis assays developed for the four distinct disorders with specific detection needs and analytical ranges could be combined on the OpenArray system and in multiplexed qPCR reactions. SNP detection of homozygous SMN1 deletions in SMA, CNV detection in the 22q11.2 critical region, and quantification of the SCID biomarker, T-cell receptor excision circles (TRECs) and CMV were all required for disease confirmation. SMA and 22q11.2 gene deletions were accurately detected using the OpenArray system, a first for the technology. The medium density deletion 22q11.2 multiplex successfully identified deletion carriers having either the larger 3 Mb deletion or the smaller 1.5 Mb deletions. Both TREC and CMV targets were detected but with a decrease in sensitivity when compared to their singleplex counterparts. Lastly, copy number detection of the TBX1 was performed when multiplexed with the TREC assay, without a decrease in detection limit of either assay. Here, we provide proof of principal that qPCR multiplexing technologies are amenable to implementation with a newborn screening laboratory.
- Published
- 2013
- Full Text
- View/download PDF
107. Drosophila Yemanuclein is required for meiosis in the oocyte and paternal chromatin assembly in the zygote
- Author
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Algazeery, Ahmed, Institut de génétique humaine (IGH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université Montpellier I, and Ounissa Aït Ahmed
- Subjects
Recombinaison ,Histone H3.3 ,Meiosis ,Oocyte ,Ovocyte ,Zygote ,Méiose ,Hira ,Recombination ,[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology - Abstract
Sexual reproduction relies on two key events: formation of cells with a haploid genome through meiosis and restoration of diploidy through syngamy in the zygote. Meiosis completion is supported exclusively by the maternal genome for the oocyte and the paternal genome for the sperm cell. In contrast diploidy restoration in the zygote is entirely dependent on maternal factors. At the end of meiosis the maternal pronucleus is competent for replication, whereas the paternal genome is packed with protamines. These proteins need to be removed in the zygote and replaced by maternally provided histones before the paternal genome acquires competence for replication, a prerequisite for syngamy. All these events must be highly coordinated to allow the first zygotic nucleus to form with the two sets of parental chromosomes and enter the first mitotic cycle. Our laboratory has identified yemanuclein-alpha, also called yemanuclein (yem) in a molecular screen for genes specifically expressed in the female germ line and its first mutant allele yem1, in a female sterile screen. The role played by yem not only in the meiotic process through which a haploid maternal pronucleus is formed but also in the zygotic process that makes a paternal pronucleus competent for syngamy, is underscored by the obtention of exceptional parthenogenetic progeny from yem1 mothers.My thesis work is precisely dedicated to the analysis of both aspects of Yemanuclein function: in the oocyte and the zygote. Using genetic, biochemical and cell biology methods we were able to uncover essential functions of Yemanuclein in early meiotic prophase in the Drosophila oocyte. Using yem1 allele (V478E), we could show its requirement for meiotic recombination especially for the frequency and timing of the double strand breaks formation. Yemanuclein association with two protein complexes, the Synaptonemal Complex (SC) and the Cohesin complex known to be required for proper chromosome segregation, supports these findings. Beyond its meiotic function, Yemanuclein is also required in the zygote for assembly of paternal pronucleus chromatin. This is achieved through a third complex that acts as histone H3.3 chaperone. In the present manuscript we identify Yemanuclein as a partner of HIRA in its role in H3.3 nucleosome assembly and deposition on the paternal pronucleus. Interestingly Yemanuclein is the first member of the HPC2/UBN1 protein family ever characterized. The role of Yem/ HPC2/ UBN1 in replication independent chromatin remodeling remained elusive until very recently. Our work is original in that it is the first to report on a role of one member of this family in oocyte meiosis and paternal chromatin assembly in the zygote.; La reproduction sexuée repose sur deux processus fondamentaux : la méiose qui permet la formation des gamètes dont le génome est haploïde et la syngamie qui permet, après fécondation, de restaurer la diploïdie par fusion des deux noyaux parentaux haploïdes. Alors que la méiose repose respectivement sur le génome maternel pour l'ovocyte et paternel pour le spermatozoïde, la restauration de la diploïdie dans le zygote repose exclusivement sur le génome maternel. Si un pronucleus maternel compétent pour la réplication est formé au terme de la méiose ovocytaire, le génome paternel quant à lui, n'acquiert cette compétence que sous l'influence de facteurs maternels. En effet, à la fin de la méiose, le génome paternel est « empaqueté » avec des protamines qui le rendent inactif pour toute fonction biologique, en particulier la réplication. L'éviction des protamines et leur remplacement par des histones maternelles sont des étapes indispensables à l'acquisition par le génome paternel de sa compétence à la réplication, préalable à la syngamie. Tous ces événements doivent être extrêmement coordonnés afin de permettre à un premier noyau zygotique comportant les deux lots de chromosomes parentaux de se former et d'entrer dans le premier cycle mitotique.Notre laboratoire a identifié yemanuclein-alpha, aussi appelé yemanuclein (yem) dans un crible moléculaire pour des gènes exprimés spécifiquement dans la lignée germinale femelle, et son premier allèle muté yem1. Cette mutation ponctuelle (V478E) a été identifiée dans un crible génétique de « stérilité femelle ». Une descendance exceptionnelle observée chez les femelles yem1, présente la propriété inattendue d'être parthénogénétique. Cette propriété révèle un double défaut chez le mutant : dans le processus de méiose ovocytaire qui conduit à la formation d'un pronucleus maternel haploïde mais aussi dans la formation d'un pronucleus paternel compétent pour la syngamie.Mes travaux de thèse ont porté sur les deux aspects de la fonction de la Yemanucléine. En conjuguant des méthodes de génétique, de biochimie, et de biologie cellulaire, nous avons pu mettre en évidence des fonctions essentielles de la Yemanucléine dans les étapes initiales de la prophase méiotique de l'ovocyte de drosophile. Nous avons pu montrer que la Yemanucléine joue un rôle clé dans la recombinaison méiotique et plus particulièrement dans la fréquence et la cinétique d'apparition des cassures double brin. Son association au complexe synaptonémal et au complexe cohésine, tous deux connus comme étant nécessaires à la ségrégation chromosomique, est un élément clé de cette fonction.Outre cette fonction méiotique, la Yemanucléine, facteur maternel, est aussi requise pour l'assemblage de la chromatine du pronucleus paternel. Nous montrons dans ce manuscrit qu'elle joue ce rôle à travers son action dans un troisième complexe, en partenariat avec la protéine HIRA. Le complexe multiprotéique contenant la protéine HIRA est connu pour sa fonction de chaperon du variant de l'histone H3.3 et son rôle dans l'assemblage de la chromatine du pronucleus paternel. La Yemanucléine est le premier membre de la famille HPC2/UBN1 caractérisé. Son rôle dans l'assemblage des nucléosomes découplé de la réplication est décrit pour la première fois dans ce manuscrit. C'est aussi la première fois qu'une protéine spécifique de la reproduction est décrite pour son implication à deux étapes clés de ce processus.
- Published
- 2013
108. La Yemanucléine de Drosophile est nécessaire à la méiose ovocytaire et l’assemblage de la chromatine paternelle dans le zygote
- Author
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Algazeery, Ahmed, Institut de génétique humaine (IGH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université Montpellier I, and Ounissa Aït Ahmed
- Subjects
Recombinaison ,Histone H3.3 ,Meiosis ,Oocyte ,Ovocyte ,Zygote ,Méiose ,Hira ,Recombination ,[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology - Abstract
Sexual reproduction relies on two key events: formation of cells with a haploid genome through meiosis and restoration of diploidy through syngamy in the zygote. Meiosis completion is supported exclusively by the maternal genome for the oocyte and the paternal genome for the sperm cell. In contrast diploidy restoration in the zygote is entirely dependent on maternal factors. At the end of meiosis the maternal pronucleus is competent for replication, whereas the paternal genome is packed with protamines. These proteins need to be removed in the zygote and replaced by maternally provided histones before the paternal genome acquires competence for replication, a prerequisite for syngamy. All these events must be highly coordinated to allow the first zygotic nucleus to form with the two sets of parental chromosomes and enter the first mitotic cycle. Our laboratory has identified yemanuclein-alpha, also called yemanuclein (yem) in a molecular screen for genes specifically expressed in the female germ line and its first mutant allele yem1, in a female sterile screen. The role played by yem not only in the meiotic process through which a haploid maternal pronucleus is formed but also in the zygotic process that makes a paternal pronucleus competent for syngamy, is underscored by the obtention of exceptional parthenogenetic progeny from yem1 mothers.My thesis work is precisely dedicated to the analysis of both aspects of Yemanuclein function: in the oocyte and the zygote. Using genetic, biochemical and cell biology methods we were able to uncover essential functions of Yemanuclein in early meiotic prophase in the Drosophila oocyte. Using yem1 allele (V478E), we could show its requirement for meiotic recombination especially for the frequency and timing of the double strand breaks formation. Yemanuclein association with two protein complexes, the Synaptonemal Complex (SC) and the Cohesin complex known to be required for proper chromosome segregation, supports these findings. Beyond its meiotic function, Yemanuclein is also required in the zygote for assembly of paternal pronucleus chromatin. This is achieved through a third complex that acts as histone H3.3 chaperone. In the present manuscript we identify Yemanuclein as a partner of HIRA in its role in H3.3 nucleosome assembly and deposition on the paternal pronucleus. Interestingly Yemanuclein is the first member of the HPC2/UBN1 protein family ever characterized. The role of Yem/ HPC2/ UBN1 in replication independent chromatin remodeling remained elusive until very recently. Our work is original in that it is the first to report on a role of one member of this family in oocyte meiosis and paternal chromatin assembly in the zygote.; La reproduction sexuée repose sur deux processus fondamentaux : la méiose qui permet la formation des gamètes dont le génome est haploïde et la syngamie qui permet, après fécondation, de restaurer la diploïdie par fusion des deux noyaux parentaux haploïdes. Alors que la méiose repose respectivement sur le génome maternel pour l'ovocyte et paternel pour le spermatozoïde, la restauration de la diploïdie dans le zygote repose exclusivement sur le génome maternel. Si un pronucleus maternel compétent pour la réplication est formé au terme de la méiose ovocytaire, le génome paternel quant à lui, n'acquiert cette compétence que sous l'influence de facteurs maternels. En effet, à la fin de la méiose, le génome paternel est « empaqueté » avec des protamines qui le rendent inactif pour toute fonction biologique, en particulier la réplication. L'éviction des protamines et leur remplacement par des histones maternelles sont des étapes indispensables à l'acquisition par le génome paternel de sa compétence à la réplication, préalable à la syngamie. Tous ces événements doivent être extrêmement coordonnés afin de permettre à un premier noyau zygotique comportant les deux lots de chromosomes parentaux de se former et d'entrer dans le premier cycle mitotique.Notre laboratoire a identifié yemanuclein-alpha, aussi appelé yemanuclein (yem) dans un crible moléculaire pour des gènes exprimés spécifiquement dans la lignée germinale femelle, et son premier allèle muté yem1. Cette mutation ponctuelle (V478E) a été identifiée dans un crible génétique de « stérilité femelle ». Une descendance exceptionnelle observée chez les femelles yem1, présente la propriété inattendue d'être parthénogénétique. Cette propriété révèle un double défaut chez le mutant : dans le processus de méiose ovocytaire qui conduit à la formation d'un pronucleus maternel haploïde mais aussi dans la formation d'un pronucleus paternel compétent pour la syngamie.Mes travaux de thèse ont porté sur les deux aspects de la fonction de la Yemanucléine. En conjuguant des méthodes de génétique, de biochimie, et de biologie cellulaire, nous avons pu mettre en évidence des fonctions essentielles de la Yemanucléine dans les étapes initiales de la prophase méiotique de l'ovocyte de drosophile. Nous avons pu montrer que la Yemanucléine joue un rôle clé dans la recombinaison méiotique et plus particulièrement dans la fréquence et la cinétique d'apparition des cassures double brin. Son association au complexe synaptonémal et au complexe cohésine, tous deux connus comme étant nécessaires à la ségrégation chromosomique, est un élément clé de cette fonction.Outre cette fonction méiotique, la Yemanucléine, facteur maternel, est aussi requise pour l'assemblage de la chromatine du pronucleus paternel. Nous montrons dans ce manuscrit qu'elle joue ce rôle à travers son action dans un troisième complexe, en partenariat avec la protéine HIRA. Le complexe multiprotéique contenant la protéine HIRA est connu pour sa fonction de chaperon du variant de l'histone H3.3 et son rôle dans l'assemblage de la chromatine du pronucleus paternel. La Yemanucléine est le premier membre de la famille HPC2/UBN1 caractérisé. Son rôle dans l'assemblage des nucléosomes découplé de la réplication est décrit pour la première fois dans ce manuscrit. C'est aussi la première fois qu'une protéine spécifique de la reproduction est décrite pour son implication à deux étapes clés de ce processus.
- Published
- 2013
109. Role of DNA damage response signalling in induction and maintenance of cellular senescence
- Author
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Pešina, František, Hodný, Zdeněk, and Janoštiak, Radoslav
- Subjects
SAHF ,p16 ,senescence ,DDR ,p21 ,HIRA ,RB ,p53 - Abstract
Cellular senescence is a state of permanent growth arrest. It is induced by many stimuli, including telomere shortening, DNA damage, oncogene hyperstimulation, chromatin perturbation and various stresses by which are cells affected. Researches showed central role of two pathways in induction and maintenance of this state. These are the p53/21 and p16/RB. The extent and dynamics of their activation by various stimuli is different. Slightly different is also their function in induction and maintenance of senescence. These differences are depicted and compared in this work.
- Published
- 2012
110. Organisation et intégrité des chromosomes parentaux à la fécondation chez la drosophile
- Author
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Orsi, Guillaume, Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Université Claude Bernard - Lyon I, Benjamin Loppin, and Pierre Couble
- Subjects
[SDV.SA]Life Sciences [q-bio]/Agricultural sciences ,Chromatine ,Dysgénésie Hybride ,Épigénétique ,Fécondation ,HIRA ,H3.3 ,Chromatin ,Hybrid dysgenesis ,Yemanuclein ,Fertilization ,Drosophila ,Epigenetics ,Yemanucléine ,Drosophile - Abstract
Sexual reproduction involves dramatic gamete differentiation and profound parental chromosomes remodelling. At fertilization, these chromosomes need to be rendered competent for the formation of the fist zygotic nucleus. I have studied the functional relevance of several paternal and maternal molecular pathways that participate during this process in Drosophila. The HIRA complex is required for nucleosome assembly in the male pronucleus at fertilization. I have further described the rôle of HIRA and its obligatory partner Yemanuclein-α during this step. I have characterized the somatic roles of this complex during nucleosome assembly and its involvment in heterochromatin stability, which gives us a better understanding of the biological needs that drive its conservation and evolution. I have also focused on several situations where parental chromosomes integrity at fertilization is compromised. (1) I have described a meiotic catastrophe associated with the natural expression of a transposon in the female germline during hybrid dysgenesis. (2) I have contributed to show that K81 is an essential protein for telomere protection in paternal chromosomes during spermiogenesis. (3) I have participated in the characterization of the chromosomal abnormalities associated with cytoplasmic incompatibility induced by Wolbachia. Together, these results underscore the specificities of parental chromosomes at fertilization and shed light into the importance of maternal and paternal pathways for their integration in the first zygotic nucleus; La reproduction sexuée implique une différentiation extrême des gamètes qui s’accompagne de profonds remaniements des chromosomes parentaux. Au moment de la fécondation, ces chromosomes doivent être rendus compétents pour la formation du premier noyau zygotique. Au cours de ma thèse, j’ai étudié l’importance fonctionnelle de plusieurs voies moléculaires paternelles et maternelles participant à cette étape chez la drosophile. Le complexe HIRA est impliqué dans l’assemblage de nucléosomes dans le pronoyau mâle à la fécondation. J’ai décrit le rôle de HIRA et de son partenaire Yemanucléine-α dans cette voie. J’ai caractérisé plus finement ce complexe en étudiant son rôle somatique dans l’assemblage des nucléosomes et son implication dans la stabilité de l’hétérochromatine, améliorant notre compréhension des besoins biologiques qui conditionnent sa conservation et son évolution. Je me suis aussi intéressé à diverses situations affectant l’intégrité des chromosomes parentaux à la fécondation. (1) J’ai décrit les conséquences catastrophiques pour la méiose femelle de l’expression naturelle d’un transposon à travers l’étude d’un cas de dysgénésie hybride. (2) J’ai contribué à montrer que la protéine K81 est essentielle pour la protection des télomères dans les chromosomes paternels au cours de la spermatogénèse. (3) J’ai participé à caractériser les conséquences pour les chromosomes paternels de l’incompatibilité cytoplasmique induite par la bactérie Wolbachia. Ensemble, ces travaux soulignent les particularités des chromosomes parentaux à la fécondation et aident à cerner l’importance des voies maternelles et paternelles dans leur intégration dans le premier noyau du zygote
- Published
- 2011
111. Organization and integrity of parental chromosomes at fertilization in Drosophila
- Author
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Orsi, Guillaume, STAR, ABES, Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Université Claude Bernard - Lyon I, Benjamin Loppin, and Pierre Couble
- Subjects
[SDV.SA]Life Sciences [q-bio]/Agricultural sciences ,[SDV.SA] Life Sciences [q-bio]/Agricultural sciences ,Chromatine ,Dysgénésie Hybride ,Épigénétique ,Fécondation ,HIRA ,H3.3 ,Hybrid dysgenesis ,Chromatin ,Yemanuclein ,Fertilization ,Epigenetics ,Drosophila ,Yemanucléine ,Drosophile - Abstract
Sexual reproduction involves dramatic gamete differentiation and profound parental chromosomes remodelling. At fertilization, these chromosomes need to be rendered competent for the formation of the fist zygotic nucleus. I have studied the functional relevance of several paternal and maternal molecular pathways that participate during this process in Drosophila. The HIRA complex is required for nucleosome assembly in the male pronucleus at fertilization. I have further described the rôle of HIRA and its obligatory partner Yemanuclein-α during this step. I have characterized the somatic roles of this complex during nucleosome assembly and its involvment in heterochromatin stability, which gives us a better understanding of the biological needs that drive its conservation and evolution. I have also focused on several situations where parental chromosomes integrity at fertilization is compromised. (1) I have described a meiotic catastrophe associated with the natural expression of a transposon in the female germline during hybrid dysgenesis. (2) I have contributed to show that K81 is an essential protein for telomere protection in paternal chromosomes during spermiogenesis. (3) I have participated in the characterization of the chromosomal abnormalities associated with cytoplasmic incompatibility induced by Wolbachia. Together, these results underscore the specificities of parental chromosomes at fertilization and shed light into the importance of maternal and paternal pathways for their integration in the first zygotic nucleus, La reproduction sexuée implique une différentiation extrême des gamètes qui s’accompagne de profonds remaniements des chromosomes parentaux. Au moment de la fécondation, ces chromosomes doivent être rendus compétents pour la formation du premier noyau zygotique. Au cours de ma thèse, j’ai étudié l’importance fonctionnelle de plusieurs voies moléculaires paternelles et maternelles participant à cette étape chez la drosophile. Le complexe HIRA est impliqué dans l’assemblage de nucléosomes dans le pronoyau mâle à la fécondation. J’ai décrit le rôle de HIRA et de son partenaire Yemanucléine-α dans cette voie. J’ai caractérisé plus finement ce complexe en étudiant son rôle somatique dans l’assemblage des nucléosomes et son implication dans la stabilité de l’hétérochromatine, améliorant notre compréhension des besoins biologiques qui conditionnent sa conservation et son évolution. Je me suis aussi intéressé à diverses situations affectant l’intégrité des chromosomes parentaux à la fécondation. (1) J’ai décrit les conséquences catastrophiques pour la méiose femelle de l’expression naturelle d’un transposon à travers l’étude d’un cas de dysgénésie hybride. (2) J’ai contribué à montrer que la protéine K81 est essentielle pour la protection des télomères dans les chromosomes paternels au cours de la spermatogénèse. (3) J’ai participé à caractériser les conséquences pour les chromosomes paternels de l’incompatibilité cytoplasmique induite par la bactérie Wolbachia. Ensemble, ces travaux soulignent les particularités des chromosomes parentaux à la fécondation et aident à cerner l’importance des voies maternelles et paternelles dans leur intégration dans le premier noyau du zygote
- Published
- 2011
112. Gearing up chromatin: A role for chromatin remodeling during the transcriptional restart upon DNA damage
- Author
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Mandemaker, I.K. (Imke), Vermeulen, W. (Wim), Marteijn, J.A. (Jurgen), Mandemaker, I.K. (Imke), Vermeulen, W. (Wim), and Marteijn, J.A. (Jurgen)
- Abstract
During transcription, RNA polymerase may encounter DNA lesions, which causes stalling of transcription. To overcome the RNA polymerase blocking lesions, the transcribed strand is repaired by a dedicated repair mechanism, called transcription coupled nucleotide excision repair (TC-NER). After repair is completed, it is essential that transcription restarts. So far, the regulation and exact molecular mechanism of this transcriptional restart upon genotoxic damage has remained elusive. Recently, three different chromatin remodeling factors, HIRA, FACT, and Dot1L, were identified to stimulate transcription restart after DNA damage. These factors either incorporate new histones or establish specific chromatin marks that will gear up the chromatin to subsequently promote transcription recovery. This adds a new layer to the current model of chromatin remodeling necessary for repair and indicates that this specific form of transcription, i.e., the transcriptional restart upon DNA damage, needs specific chromatin remodeling events.
- Published
- 2014
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113. HIRA, a conserved histone chaperone, plays an essential role in low-dose stress response via transcriptional stimulation in fission yeast.
- Author
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70551381, 60143369, 30184493, Chujo, Moeko, Tarumoto, Yusuke, Miyatake, Koichi, Nishida, Eisuke, Ishikawa, Fuyuki, 70551381, 60143369, 30184493, Chujo, Moeko, Tarumoto, Yusuke, Miyatake, Koichi, Nishida, Eisuke, and Ishikawa, Fuyuki
- Published
- 2012
114. HIRA deficiency in muscle fibers causes hypertrophy and susceptibility to oxidative stress.
- Author
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Valenzuela, Nicolas, Soibam, Benjamin, Lerong Li, Jing Wang, Byers, Lauren A., Yu Liu, Schwartz, Robert J., and Stewart, M. David
- Subjects
- *
CHROMATIN , *DNA replication , *CELL cycle , *LABORATORY mice , *MUSCLE growth , *SKELETAL muscle - Abstract
Nucleosome assembly proceeds through DNA replication-coupled or replication-independent mechanisms. For skeletal myocytes, whose nuclei have permanently exited the cell cycle, replication-independent assembly is the only mode available for chromatin remodeling. For this reason, any nucleosome composition alterations accompanying transcriptional responses to physiological signals must occur through a DNA replication-independent pathway. HIRA is the histone chaperone primarily responsible for replication-independent incorporation of histone variant H3.3 across gene bodies and regulatory regions. Thus, HIRA would be expected to play an important role in epigenetically regulating myocyte gene expression. The objective of this study was to determine the consequence of eliminating HIRA from mouse skeletal myocytes. At 6 weeks of age, myofibers lacking HIRA showed no pathological abnormalities; however, genes involved in transcriptional regulation were downregulated. By 6 months of age, myofibers lacking HIRA exhibited hypertrophy, sarcolemmal perforation and oxidative damage. Genes involved in muscle growth and development were upregulated, but those associated with responses to cellular stresses were downregulated. These data suggest that elimination of HIRA produces a hypertrophic response in skeletal muscle and leaves myofibers susceptible to stress-induced degeneration. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
115. Nationwide pulmonary function test rates in South Korean asthma patients.
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Choi JY, Yoon HK, Lee JH, Yoo KH, Kim BY, Bae HW, Kim YK, and Rhee CK
- Abstract
Background: Previous studies have shown that pulmonary function tests (PFTs) are performed at considerably lower rates than would be expected if standard guidelines were followed. The goal of this study was to evaluate the current status of PFT performance in the Republic of Korea., Methods: We analysed quality assessment data from a nationwide Health Insurance Review and Assessment Service database collected from July 2013 to June 2014. PFT performance rates were compared among types and specialties of medical institutions. PFT performance rates were also measured by patient gender, age, and insurance type. Possession rates of PFT equipment and performance rates of each type of PFT were also evaluated., Results: A total of 16,804 institutions and 831,613 patients were included in this study. The mean overall PFT performance rate was 22.67%. The performance rate in tertiary hospitals was 78.00%, while PFTs were performed in only 20.87% of asthma patients at primary health clinics. Male and elderly patients received PFTs more frequently than did female and younger patients. Also, patients who were covered by the Korean Veterans Health Service received a PFT more frequently than those covered by other insurance services. The possession rate of PFT equipment was significantly higher in tertiary hospitals than in primary health clinics. Of all PFT types, spirometry with flow-volume curve was performed for most patients., Conclusions: The PFT performance rate was significantly lower than would be expected if guidelines were followed. Average performance rates were higher in tertiary hospitals and for male and elderly patients., Competing Interests: Conflicts of Interest: The authors have no conflicts of interest to declare.
- Published
- 2018
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116. Differential Expression of Histone H3.3 Genes and Their Role in Modulating Temperature Stress Response in Caenorhabditis elegans .
- Author
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Delaney K, Mailler J, Wenda JM, Gabus C, and Steiner FA
- Subjects
- Animals, Caenorhabditis elegans growth & development, Caenorhabditis elegans metabolism, Caenorhabditis elegans Proteins genetics, Germ Cells cytology, Germ Cells metabolism, Histone Chaperones genetics, Histone Chaperones metabolism, Histones genetics, Infertility genetics, Nucleosomes metabolism, Caenorhabditis elegans genetics, Caenorhabditis elegans Proteins metabolism, Heat-Shock Response, Histones metabolism
- Abstract
Replication-independent variant histones replace canonical histones in nucleosomes and act as important regulators of chromatin function. H3.3 is a major variant of histone H3 that is remarkably conserved across taxa and is distinguished from canonical H3 by just four key amino acids. Most genomes contain two or more genes expressing H3.3, and complete loss of the protein usually causes sterility or embryonic lethality. Here, we investigate the developmental expression patterns of the five Caenorhabditis elegans H3.3 homologs and identify two previously uncharacterized homologs to be restricted to the germ line. Despite these specific expression patterns, we find that neither loss of individual H3.3 homologs nor the knockout of all five H3.3-coding genes causes sterility or lethality. However, we demonstrate an essential role for the conserved histone chaperone HIRA in the nucleosomal loading of all H3.3 variants. This requirement can be bypassed by mutation of the H3.3-specific residues to those found in H3. While even removal of all H3.3 homologs does not result in lethality, it leads to reduced fertility and viability in response to high-temperature stress. Thus, our results show that H3.3 is nonessential in C. elegans but is critical for ensuring adequate response to stress., (Copyright © 2018 by the Genetics Society of America.)
- Published
- 2018
- Full Text
- View/download PDF
117. Characterization of H3.3 and HIRA expression and function in bovine early embryos.
- Author
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Zhang K, Wang H, Rajput SK, Folger JK, and Smith GW
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- Animals, Cell Lineage genetics, Embryo, Mammalian, Embryonic Development genetics, Female, Gene Expression Regulation, Developmental, Gene Knockdown Techniques, Histone Chaperones metabolism, Histones metabolism, Male, Pregnancy, Zygote metabolism, Blastocyst metabolism, Cattle embryology, Cattle genetics, Histone Chaperones genetics, Histone Chaperones physiology, Histones genetics
- Abstract
Histone variant H3.3 is encoded by two distinct genes, H3F3A and H3F3B, that are closely associated with actively transcribed genes. H3.3 replacement is continuous and essential for maintaining correct chromatin structure during mouse oogenesis. Upon fertilization, H3.3 is incorporated to parental chromatin, and is required for blastocyst formation in mice. The H3.3 exchange process is facilitated by the chaperone HIRA, particularly during zygote development. We previously demonstrated that H3.3 is required for bovine early embryonic development; here, we explored the mechanisms of its functional requirement. H3F3A mRNA abundance is stable whereas H3F3B and HIRA mRNA are relatively dynamic during early embryonic development. H3F3B mRNA quantity is also considerably higher than H3F3A. Immunofluorescence analysis revealed an even distribution of H3.3 between paternal and maternal pronuclei in zygotes, and subsequent stage-specific localization of H3.3 in early bovine embryos. Knockdown of H3.3 by targeting both H3F3A and H3F3B dramatically decreased the expression of NANOG (a pluripotency marker) and CTGF (Connective tissue growth factor; a trophectoderm marker) in bovine blastocysts. Additionally, we noted that Histone H3 lysine 36 dimethylation and linker Histone H1 abundance is reduced in H3.3-deficient embryos, which was similar to effects following knockdown of CHD1 (Chromodomain helicase DNA-binding protein 1). By contrast, no difference was observed in the abundance of Histone H3 lysine 4 trimethylation, Histone H3 lysine 9 dimethylation, or Splicing factor 3 B1. Collectively, these results established that H3.3 is required for correct epigenetic modifications and H1 deposition, dysregulation of which likely mediate the poor development in H3.3-deficient embryos., (© 2017 Wiley Periodicals, Inc.)
- Published
- 2018
- Full Text
- View/download PDF
118. Imaging Newly Synthesized and Old Histone Variant Dynamics Dependent on Chaperones Using the SNAP-Tag System.
- Author
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Torné J, Orsi GA, Ray-Gallet D, and Almouzni G
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- HeLa Cells, Humans, Protein Isoforms biosynthesis, Staining and Labeling, Histone Chaperones metabolism, Histones biosynthesis, Imaging, Three-Dimensional, Molecular Biology methods
- Abstract
Distinct histone variants mark chromatin domains in the nucleus. To understand how these marks are established and maintained, one has to decipher how the dynamic distribution of these variants is orchestrated. These dynamics are associated with all DNA-based processes such as DNA replication, repair, transcription, heterochromatin formation and chromosome segregation. Key factors, known as histone chaperones, have been involved in escorting histones, thereby contributing to the chromatin landscape of given cell types. SNAP-tag-based imaging system enables the distinction between old and newly deposited histones, and has proved to be a powerful method for the visualization of histone variant dynamics on a cell-by-cell basis. This approach enables the tracking of specific variants in vivo and defining their timing and mode of deposition throughout the cell cycle and in different nuclear territories. Here, we provide a detailed protocol to exploit the SNAP-tag technology to assess the dynamics of newly synthesized and old histones. We then show that combining the SNAP-tagging of histones with the knockdown of candidate factors, represents an effective approach to decipher the role of key actors in guiding histone dynamics. Here, we specifically illustrate how this strategy was used to identify the essential role of the chaperone HIRA in deposition of newly synthesized histone variant H3.3.
- Published
- 2018
- Full Text
- View/download PDF
119. PENERAPAN SISTEM MANAJEMEN KESELAMATAN DAN KESEHATAN KERJA (K3) DAN IDENTIFIKASI POTENSI BAHAYA KERJA (Studi kasus di PT. LTX Kota Cilegon- Banten)
- Author
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Feni Akbar Rini and Wahyu Susihono
- Subjects
lcsh:T55.4-60.8 ,lcsh:Industrial engineering. Management engineering ,lcsh:Industry ,Potensi bahaya ,lcsh:HD2321-4730.9 ,Penerapan SMK3 ,HIRA ,General Medicine ,FTA - Abstract
Perlindungan terhadap keselamatan dan kesehatan kerja masih jauh dari yang diharapkan karena masih banyak terjadi kecelakaan kerja serta potensi bahaya kerja yang dapat membahayakan tenaga kerja. Penerapkan sistem manajeman keselamatan dan kesehatan kerja (SMK3) perlu dilakukan secara optimal. Penerapan SMK3 di perusahaan belum tentu berbanding lurus terhadap potensi bahaya (hazard) yang ada di lingkungan sekitar perusahaan. Penelitian ini bertujuan untuk mengetahui nilai risiko potensi bahaya kerja dan kategori potensi bahaya kerja di perusahaan serta mengetahui faktor penyebab terbesar terjadinya kecelakaan kerja di perusahaan. Penelitian ini menggunakan pendekatan metode HIRA dan FTA. Hasil yang diperoleh menunjukkan bahwa penerapan SMK3 telah sesuai dengan undang-Undang yang berlaku, namun nilai resiko potensi bahaya bagian fluid utility menunjukkan tingkat keparahan bahaya kerja kecil dan kemungkinan terjadinya potensi bahaya kerja juga kecil, nilai kategori potensi bahaya kerja perlu dikendalikan dengan prosedur rutin. Faktor penyebab potensial terjadinya potensi bahaya adalah suara mesin bising, Standard Operational procedure (SOP) belum terpasang secara ergonomis, terdapat benda asing yang menghalangi jalan, temperatur ruangan meningkat 50C dari temperatur normal. Kata Kunci : Penerapan SMK3, Potensi bahaya, HIRA, FTA
- Published
- 2013
- Full Text
- View/download PDF
120. Quelques précisions sur le matériel de Hira: (céramique et verre)
- Author
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Marie-Odile Rousset, Histoire, Archéologie et Littératures des mondes chrétiens et musulmans médiévaux (CIHAM), École normale supérieure - Lyon (ENS Lyon)-Université Lumière - Lyon 2 (UL2)-École des hautes études en sciences sociales (EHESS)-Université Jean Moulin - Lyon 3 (UJML), Université de Lyon-Université de Lyon-Avignon Université (AU)-Centre National de la Recherche Scientifique (CNRS), ORIENT ET MÉDITERRANÉE : Textes, Archéologie, Histoire (OM), Université Paris 1 Panthéon-Sorbonne (UP1)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure de Lyon (ENS de Lyon)-Université Lumière - Lyon 2 (UL2)-École des hautes études en sciences sociales (EHESS)-Université Jean Moulin - Lyon 3 (UJML), and Université Paris 1 Panthéon-Sorbonne (UP1)-École Pratique des Hautes Études (EPHE)
- Subjects
verre ,atelier ,workshop ,[SHS.ARCHEO]Humanities and Social Sciences/Archaeology and Prehistory ,archéologie ,Iraq ,archaeology ,pottery ,Kufa ,céramique ,Hira ,glass - Abstract
The aim of this article is to describe some pottery and glass cherds from Hira, in Iraq, who was the lakhmid dynastie's capital before Islam. The pieces were collected by a french archaeological mission in Iraq in 1990. Some material, found by David Talbot-Rice in the 1930's and stored in the Ashmolean Museum of Oxford were also analysed. There was at Hira a workshop producing moulded, incised and barbotine applied wares in the 7th - 8th c.; Le but de cet article est de décrire des tessons de céramique et de verre provenant de Hira, en Iraq, qui fut la capitale de la dynastie lakhmide durant l'époque précédant l'arrivée de l'Islam. Les pièces ont été collectée par une mission archéologique française, en 1990. Le matériel trouvé par David Talbot-Rice dans les années trente et conservé à l'Ashmoleum Museum d'Oxford a également été analysé. Il y avait à Hira un atelier qui produisait des céramiques à décor moulé, incisé et appliqué, aux 7e - 8e siècles.
- Published
- 1994
121. Quelques précisions sur le matériel de Hira
- Author
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Rousset, Marie-Odile and Rousset, Marie-Odile
- Subjects
verre ,atelier ,workshop ,[SHS.ARCHEO] Humanities and Social Sciences/Archaeology and Prehistory ,archéologie ,Iraq ,archaeology ,pottery ,Kufa ,céramique ,Hira ,glass - Abstract
The aim of this article is to describe some pottery and glass cherds from Hira, in Iraq, who was the lakhmid dynastie's capital before Islam. The pieces were collected by a french archaeological mission in Iraq in 1990. Some material, found by David Talbot-Rice in the 1930's and stored in the Ashmolean Museum of Oxford were also analysed. There was at Hira a workshop producing moulded, incised and barbotine applied wares in the 7th - 8th c., Le but de cet article est de décrire des tessons de céramique et de verre provenant de Hira, en Iraq, qui fut la capitale de la dynastie lakhmide durant l'époque précédant l'arrivée de l'Islam. Les pièces ont été collectée par une mission archéologique française, en 1990. Le matériel trouvé par David Talbot-Rice dans les années trente et conservé à l'Ashmoleum Museum d'Oxford a également été analysé. Il y avait à Hira un atelier qui produisait des céramiques à décor moulé, incisé et appliqué, aux 7e - 8e siècles.
- Published
- 1994
122. La céramique de Hira à décor moulé, incisé ou appliqué. Techniques de fabrication et aperçu de la diffusion
- Author
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Marie-Odile Rousset, Institut Français d'Études Arabes de Damas (IFEAD), Ministère de l'Europe et des Affaires étrangères (MEAE), ORIENT ET MÉDITERRANÉE : Textes, Archéologie, Histoire (OM), Université Paris 1 Panthéon-Sorbonne (UP1)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), E. Villeneuve et P. M. Watson, and Rousset, Marie-Odile
- Subjects
atelier ,[SHS.ARCHEO] Humanities and Social Sciences/Archaeology and Prehistory ,workshop ,[SHS.ARCHEO]Humanities and Social Sciences/Archaeology and Prehistory ,Iraq ,pottery ,Hira ,céramique - Abstract
International audience; The aim of this article is to describe some pottery cherds from Hira, in Iraq. The pieces were collected by a french archaeological mission in Iraq in 1990. Some material, found by David Talbot-Rice in the 1930's and stored in the Ashmolean Museum of Oxford were also analysed. There was at Hira a workshop producing moulded, incised and barbotine applied wares in the 7th - 8th c.v
- Published
- 1994
123. ChIP-Sequencing to Map the Epigenome of Senescent Cells Using Benzonase Endonuclease.
- Author
-
Rai TS and Adams PD
- Subjects
- Animals, Cell Cycle Checkpoints, Cellular Senescence, Chromatin genetics, Chromatin metabolism, DNA genetics, DNA metabolism, Humans, Sequence Analysis, DNA methods, Chromatin Immunoprecipitation methods, Endodeoxyribonucleases metabolism, Endoribonucleases metabolism, Epigenomics methods, Serratia marcescens enzymology
- Abstract
Cellular senescence is a state of stable cell cycle arrest triggered by diverse stresses. Establishment of senescence occurs in conjunction with a multitude of chromatin changes, which are just beginning to be studied. These chromatin changes are hypothesized to be causative for senescence. Currently, a preferred method to study such changes is chromatin immunoprecipitation followed by sequencing (ChIP-Seq). This is usually done by cross-linking the cells with formaldehyde and then generating chromatin fragments between 150 and 300bp by sonication. The DNA replication-independent histone chaperone HIRA plays an important role in control of chromatin in nonproliferating senescent cells. While investigating the role of HIRA in senescence, we found conventional ChIP protocols to be problematic, routinely yielding too low amounts of DNA for sequencing. To overcome these problems we adapted and optimized an alternative ChIP method that does not rely on cross-linking and sonication for chromatin fragmentation, and is able to easily isolate chromatin from senescent cells ready for immunoprecipitation. This method uses Benzonase endonuclease for solubilization of uncross-linked chromatin by digestion of DNA and RNA, in the absence of proteolytic activity. Using this protocol, we were easily able to immunoprecipitate HIRA with sufficient DNA for Illumina sequencing., (© 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
124. Functional Characterization of Histone Chaperones Using SNAP-Tag-Based Imaging to Assess De Novo Histone Deposition.
- Author
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Clément C, Vassias I, Ray-Gallet D, and Almouzni G
- Subjects
- Animals, Cell Culture Techniques methods, Chromatin metabolism, DNA Damage, Fluorescent Dyes analysis, Fluorescent Dyes metabolism, Guanine analogs & derivatives, Guanine metabolism, Histone Chaperones analysis, Histone Chaperones genetics, Histones analysis, Histones genetics, Humans, Microscopy, Fluorescence methods, Mutation, O(6)-Methylguanine-DNA Methyltransferase analysis, O(6)-Methylguanine-DNA Methyltransferase metabolism, Optical Imaging methods, Recombinant Fusion Proteins analysis, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Histone Chaperones metabolism, Histones metabolism, O(6)-Methylguanine-DNA Methyltransferase genetics
- Abstract
Histone chaperones-key actors in the dynamic organization of chromatin-interact with the various histone variants to ensure their transfer in and out of chromatin. In vitro chromatin assembly assays and isolation of protein complexes using tagged histone variants provided first clues concerning their binding specificities and mode of action. Here, we describe an in vivo method using SNAP-tag-based imaging to assess the de novo deposition of histones and the role of histone chaperones. This method exploits cells expressing SNAP-tagged histones combined with individual cell imaging to visualize directly de novo histone deposition in vivo. We show how, by combining this method with siRNA-based depletion, we could assess the function of two distinct histone chaperones. For this, we provide the details of the method as applied in two examples to characterize the function of the histone chaperones CAF-1 and HIRA. In both cases, we document the impact of their depletion on the de novo deposition of the histone variants H3.1 and H3.3, first in a normal context and second in response to DNA damage. We discuss how this cellular assay offers means to define in a systematic manner the function of any chosen chaperone with respect to the deposition of a given histone variant., (© 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
125. Phosphorylation of H4 Ser 47 promotes HIRA-mediated nucleosome assembly.
- Author
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Bin Kang, Mintie Pu, Gangqing Hu, Weihong Wen, Zigang Dong, Keji Zhao, Stillman, Bruce, and Zhiguo Zhan
- Subjects
- *
HISTONES , *PHOSPHORYLATION , *CHROMATIN , *PROTEIN kinases , *CATALYSIS - Abstract
Histone H3 variant H3.3, while differing from canonical H3 (H3.1) by only five amino acids, is assembled into nucleosomes, along with histone H4, at genic regions by the histone chaperone HIRA, whereas H3.1 is assembled into nucleosomes in a CAF-1-dependent reaction. Here, we show that phosphorylation of histone H4 Ser 47 (H4S47ph), catalyzed by the PAK2 kinase, promotes nucleosome assembly of H3.3-H4 and inhibits nucleosome assembly of H3.1-H4 by increasing the binding affinity of HIRA to H3.3-H4 and reducing association of CAF-1 with H3.1-H4. These results reveal a mechanism whereby H4S47ph distinctly regulates nucleosome assembly of H3.1 and H3.3. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
126. Natural course of early COPD
- Author
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Chin Kook Rhee, Kim K, Hk, Yoon, Ja, Kim, Sh, Kim, Sh, Lee, Yb, Park, Ks, Jung, Kh, Yoo, and Yi, Hwang
- Subjects
lcsh:RC705-779 ,early COPD ,NHI ,cost ,utilization ,HIRA ,lcsh:Diseases of the respiratory system ,KNAHNES - Abstract
Chin Kook Rhee,1 Kyungjoo Kim,1 Hyoung Kyu Yoon,2 Jee-Ae Kim,3 Sang Hyun Kim,4 Sang Haak Lee,5 Yong Bum Park,6 Ki-Suck Jung,7 Kwang Ha Yoo,8 Yong Il Hwang7 1Division of Pulmonary, Allergy and Critical Care Medicine, Department of Internal Medicine, Seoul St Mary’s Hospital, 2Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Yeouido St Mary’s Hospital, College of Medicine, The Catholic University of Korea, Seoul, 3Pharmaceutical Policy Evaluation Research Team, Research Institution, 4Big Data Division, Health Insurance Review and Assessment Service, Wonju, 5Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, St Paul’s Hospital, College of Medicine, The Catholic University of Korea, 6Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Internal Medicine, Hallym University Kangdong Sacred Heart Hospital, Seoul, 7Division of Pulmonary, Allergy and Critical Care Medicine, Department of Internal Medicine, Hallym University Sacred Heart Hospital, Hallym University College of Medicine, Anyang, 8Division of Pulmonary, Allergy and Critical Care Medicine, Department of Internal Medicine, Konkuk University School of Medicine, Seoul, Republic of Korea Background and objective: Few studies have examined the natural course of early COPD. The aim of this study was to observe the natural course of early COPD patients. We also aimed to analyze medical utilization and costs for early COPD during a 6-year period. Methods: Patients with early COPD were selected from Korean National Health and Nutrition Examination Survey (KNHANES) data. We linked the KNHANES data of patients with early COPD to National Health Insurance data. Results: A total of 2,397 patients were enrolled between 2007 and 2012. The mean forced expiratory volume in 1second (FEV1) was 78.6%, and the EuroQol five dimensions questionnaire (EQ-5D) index value was 0.9. In total, 110 patients utilized health care for COPD in 2007, and this number increased to 179 in 2012. The total mean number of days used per person increased from 4.9 in 2007 to 7.8 in 2012. The total medical cost per person also increased from 248.8 US dollar (USD) in 2007 to 780.6 USD in 2013. A multiple linear regression revealed that age, lower body mass index, lower FEV1 (%), and lower EQ-5D score were significantly associated with medical costs. Conclusion: Even in early COPD patients, some of them eventually progressed and utilized health care for COPD. Keywords: early COPD, KNHANES, NHI, HIRA, utilization, cost
127. O-linked N -acetylglucosamine transferase (OGT) interacts with the histone chaperone HIRA complex and regulates nucleosome assembly and cellular senescence
- Author
-
Lee, Jong-Sun and Zhang, Zhiguo
- Published
- 2016
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